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Yorodumi- EMDB-25037: Cryo-EM 3D map of the human Exostosin-1 and Exostosin-2 heterodim... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-25037 | |||||||||
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Title | Cryo-EM 3D map of the human Exostosin-1 and Exostosin-2 heterodimer in complex with a 7-sugar oligosaccharide acceptor analog | |||||||||
Map data | ||||||||||
Sample |
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Function / homology | Function and homology information glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase / N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase / hypersensitivity / heart field specification / lymphocyte adhesion to endothelial cell of high endothelial venule / heparan sulfate N-acetylglucosaminyltransferase activity / glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase activity / N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase activity / smoothened signaling pathway involved in lung development / developmental growth involved in morphogenesis ...glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase / N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase / hypersensitivity / heart field specification / lymphocyte adhesion to endothelial cell of high endothelial venule / heparan sulfate N-acetylglucosaminyltransferase activity / glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase activity / N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase activity / smoothened signaling pathway involved in lung development / developmental growth involved in morphogenesis / sweat gland development / perichondral bone morphogenesis / mesenchymal cell differentiation involved in bone development / response to leukemia inhibitory factor / chondroitin sulfate metabolic process / UDP-N-acetylglucosamine transferase complex / response to heparin / chondrocyte hypertrophy / embryonic skeletal joint development / hematopoietic stem cell migration to bone marrow / : / heparan sulfate proteoglycan biosynthetic process, polysaccharide chain biosynthetic process / fluid transport / glucuronosyltransferase activity / limb joint morphogenesis / tight junction organization / heparin biosynthetic process / Defective EXT2 causes exostoses 2 / Defective EXT1 causes exostoses 1, TRPS2 and CHDS / stomach development / sebaceous gland development / glomerular basement membrane development / dendrite self-avoidance / heparan sulfate proteoglycan biosynthetic process / HS-GAG biosynthesis / glycosaminoglycan biosynthetic process / lymphocyte migration into lymphoid organs / chondrocyte proliferation / sulfation / hematopoietic stem cell homeostasis / podocyte differentiation / glandular epithelial cell differentiation / dendritic cell migration / endochondral bone morphogenesis / polysaccharide biosynthetic process / sodium ion homeostasis / endochondral bone growth / acetylglucosaminyltransferase activity / response to light intensity / basement membrane organization / cartilage development involved in endochondral bone morphogenesis / cranial skeletal system development / vocalization behavior / stem cell division / endoderm development / multicellular organismal-level water homeostasis / leukocyte tethering or rolling / vacuole organization / olfactory bulb development / endochondral ossification / fear response / optic nerve development / protein N-linked glycosylation / cellular response to fibroblast growth factor stimulus / neural crest cell differentiation / collagen fibril organization / ossification involved in bone maturation / motor behavior / cell adhesion mediated by integrin / epithelial tube branching involved in lung morphogenesis / hair follicle morphogenesis / heart contraction / glycosyltransferase activity / protein glycosylation / social behavior / mesoderm development / mesoderm formation / catalytic complex / antigen processing and presentation / canonical Wnt signaling pathway / gastrulation / hematopoietic stem cell differentiation / fibroblast growth factor receptor signaling pathway / blood vessel remodeling / cell fate commitment / chondrocyte differentiation / BMP signaling pathway / bone resorption / ossification / synaptic transmission, glutamatergic / axon guidance / protein catabolic process / multicellular organism growth / wound healing / cellular response to virus / regulation of blood pressure / vasodilation / gene expression / protein-containing complex assembly / protein heterodimerization activity Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.8 Å | |||||||||
Authors | Li H | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Nat Chem Biol / Year: 2023 Title: Structural basis for heparan sulfate co-polymerase action by the EXT1-2 complex. Authors: Hua Li / Digantkumar Chapla / Robert A Amos / Annapoorani Ramiah / Kelley W Moremen / Huilin Li / Abstract: Heparan sulfate (HS) proteoglycans are extended (-GlcAβ1,4GlcNAcα1,4-) co-polymers containing decorations of sulfation and epimerization that are linked to cell surface and extracellular matrix ...Heparan sulfate (HS) proteoglycans are extended (-GlcAβ1,4GlcNAcα1,4-) co-polymers containing decorations of sulfation and epimerization that are linked to cell surface and extracellular matrix proteins. In mammals, HS repeat units are extended by an obligate heterocomplex of two exostosin family members, EXT1 and EXT2, where each protein monomer contains distinct GT47 (GT-B fold) and GT64 (GT-A fold) glycosyltransferase domains. In this study, we generated human EXT1-EXT2 (EXT1-2) as a functional heterocomplex and determined its structure in the presence of bound donor and acceptor substrates. Structural data and enzyme activity of catalytic site mutants demonstrate that only two of the four glycosyltransferase domains are major contributors to co-polymer syntheses: the EXT1 GT-B fold β1,4GlcA transferase domain and the EXT2 GT-A fold α1,4GlcNAc transferase domain. The two catalytic sites are over 90 Å apart, indicating that HS is synthesized by a dissociative process that involves a single catalytic site on each monomer. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_25037.map.gz | 56.8 MB | EMDB map data format | |
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Header (meta data) | emd-25037-v30.xml emd-25037.xml | 21.9 KB 21.9 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_25037_fsc.xml | 9.2 KB | Display | FSC data file |
Images | emd_25037.png | 162.6 KB | ||
Masks | emd_25037_msk_1.map | 64 MB | Mask map | |
Others | emd_25037_additional_1.map.gz emd_25037_half_map_1.map.gz emd_25037_half_map_2.map.gz | 6.4 MB 49.7 MB 49.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-25037 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-25037 | HTTPS FTP |
-Related structure data
Related structure data | 7sckMC 7schC 7scjC 7uqxC 7uqyC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_25037.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.828 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_25037_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: #1
File | emd_25037_additional_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_25037_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_25037_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : human EXT1/2
Entire | Name: human EXT1/2 |
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Components |
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-Supramolecule #1: human EXT1/2
Supramolecule | Name: human EXT1/2 / type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: #1-#2 |
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Source (natural) | Organism: Homo sapiens (human) |
-Macromolecule #1: Exostosin-1
Macromolecule | Name: Exostosin-1 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO EC number: glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 83.404023 KDa |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: GFRASRSHSR REEHSGRNGL HHPSPDHFWP RFPDALRPFV PWDQLENEDS SVHISPRQKR DANSSIYKGK KCRMESCFDF TLCKKNGFK VYVYPQQKGE KIAESYQNIL AAIEGSRFYT SDPSQACLFV LSLDTLDRDQ LSPQYVHNLR SKVQSLHLWN N GRNHLIFN ...String: GFRASRSHSR REEHSGRNGL HHPSPDHFWP RFPDALRPFV PWDQLENEDS SVHISPRQKR DANSSIYKGK KCRMESCFDF TLCKKNGFK VYVYPQQKGE KIAESYQNIL AAIEGSRFYT SDPSQACLFV LSLDTLDRDQ LSPQYVHNLR SKVQSLHLWN N GRNHLIFN LYSGTWPDYT EDVGFDIGQA MLAKASISTE NFRPNFDVSI PLFSKDHPRT GGERGFLKFN TIPPLRKYML VF KGKRYLT GIGSDTRNAL YHVHNGEDVV LLTTCKHGKD WQKHKDSRCD RDNTEYEKYD YREMLHNATF CLVPRGRRLG SFR FLEALQ AACVPVMLSN GWELPFSEVI NWNQAAVIGD ERLLLQIPST IRSIHQDKIL ALRQQTQFLW EAYFSSVEKI VLTT LEIIQ DRIFKHISRN SLIWNKHPGG LFVLPQYSSY LGDFPYYYAN LGLKPPSKFT AVIHAVTPLV SQSQPVLKLL VAAAK SQYC AQIIVLWNCD KPLPAKHRWP ATAVPVVVIE GESKVMSSRF LPYDNIITDA VLSLDEDTVL STTEVDFAFT VWQSFP ERI VGYPARSHFW DNSKERWGYT SKWTNDYSMV LTGAAIYHKY YHYLYSHYLP ASLKNMVDQL ANCEDILMNF LVSAVTK LP PIKVTQKKQY KETMMGQTSR ASRWADPDHF AQRQSCMNTF ASWFGYMPLI HSQMRLDPVL FKDQVSILRK KYRDIERL |
-Macromolecule #2: Exostosin-2
Macromolecule | Name: Exostosin-2 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO EC number: glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 77.238031 KDa |
Recombinant expression | Organism: Homo sapiens (human) |
Sequence | String: GWPHSIESSN DWNVEKRSIR DVPVVRLPAD SPIPERGDLS CRMHTCFDVY RCGFNPKNKI KVYIYALKKY VDDFGVSVSN TISREYNEL LMAISDSDYY TDDINRACLF VPSIDVLNQN TLRIKETAQA MAQLSRWDRG TNHLLFNMLP GGPPDYNTAL D VPRDRALL ...String: GWPHSIESSN DWNVEKRSIR DVPVVRLPAD SPIPERGDLS CRMHTCFDVY RCGFNPKNKI KVYIYALKKY VDDFGVSVSN TISREYNEL LMAISDSDYY TDDINRACLF VPSIDVLNQN TLRIKETAQA MAQLSRWDRG TNHLLFNMLP GGPPDYNTAL D VPRDRALL AGGGFSTWTY RQGYDVSIPV YSPLSAEVDL PEKGPGPRQY FLLSSQVGLH PEYREDLEAL QVKHGESVLV LD KCTNLSE GVLSVRKRCH KHQVFDYPQV LQEATFCVVL RGARLGQAVL SDVLQAGCVP VVIADSYILP FSEVLDWKRA SVV VPEEKM SDVYSILQSI PQRQIEEMQR QARWFWEAYF QSIKAIALAT LQIINDRIYP YAAISYEEWN DPPAVKWGSV SNPL FLPLI PPQSQGFTAI VLTYDRVESL FRVITEVSKV PSLSKLLVVW NNQNKNPPED SLWPKIRVPL KVVRTAENKL SNRFF PYDE IETEAVLAID DDIIMLTSDE LQFGYEVWRE FPDRLVGYPG RLHLWDHEMN KWKYESEWTN EVSMVLTGAA FYHKYF NYL YTYKMPGDIK NWVDAHMNCE DIAMNFLVAN VTGKAVIKVT PRKKFKCPEC TAIDGLSLDQ THMVERSECI NKFASVF GT MPLKVVEHRA DPVLYKDDFP EKLKSFPNIG SL |
-Macromolecule #5: URIDINE-5'-DIPHOSPHATE
Macromolecule | Name: URIDINE-5'-DIPHOSPHATE / type: ligand / ID: 5 / Number of copies: 2 / Formula: UDP |
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Molecular weight | Theoretical: 404.161 Da |
Chemical component information | ChemComp-UDP: |
-Macromolecule #6: 2-acetamido-2-deoxy-beta-D-glucopyranose
Macromolecule | Name: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 6 / Number of copies: 1 / Formula: NAG |
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Molecular weight | Theoretical: 221.208 Da |
Chemical component information | ChemComp-NAG: |
-Macromolecule #7: MANGANESE (II) ION
Macromolecule | Name: MANGANESE (II) ION / type: ligand / ID: 7 / Number of copies: 1 / Formula: MN |
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Molecular weight | Theoretical: 54.938 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.3 mg/mL | |||||||||
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Buffer | pH: 7 Component:
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Grid | Model: Quantifoil R2/1 / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 299 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.9 µm / Nominal magnification: 105000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Temperature | Min: 193.0 K / Max: 193.0 K |
Alignment procedure | Coma free - Residual tilt: 0.05 mrad |
Image recording | #0 - Image recording ID: 1 / #0 - Film or detector model: GATAN K3 (6k x 4k) / #0 - Number grids imaged: 1 / #0 - Number real images: 5108 / #0 - Average exposure time: 1.5 sec. / #0 - Average electron dose: 68.0 e/Å2 / #1 - Image recording ID: 2 / #1 - Film or detector model: GATAN K3 (6k x 4k) / #1 - Digitization - Dimensions - Width: 5760 pixel / #1 - Digitization - Dimensions - Height: 4092 pixel / #1 - Number grids imaged: 1 / #1 - Number real images: 7609 / #1 - Average exposure time: 1.5 sec. / #1 - Average electron dose: 72.0 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
-Atomic model buiding 1
Refinement | Protocol: AB INITIO MODEL |
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Output model | PDB-7sck: |